title: Development and preclinical characterization of a humanized antibody targeting CXCL12.
abstract: OBJECTIVE
  Our goal was to develop a potent humanized antibody against mouse/human CXCL12. This report summarized its in vitro and in vivo activities.
  
  
  METHODS
  Cell surface binding and cell migration assays were used to select neutralizing hamster antibodies, followed by testing in several animal models. Monoclonal antibody (mAb) 30D8 was selected for humanization based on its in vitro and in vivo activities.
  
  
  RESULTS
  30D8, a hamster antibody against mouse and human CXCL12α, CXCL12β, and CXCL12γ, was shown to dose-dependently block CXCL12α binding to CXCR4 and CXCR7, and CXCL12α-induced Jurkat cell migration in vitro. Inhibition of primary tumor growth and/or metastasis was observed in several models. 30D8 alone significantly ameliorated arthritis in a mouse collagen-induced arthritis model (CIA). Combination with a TNF-α antagonist was additive. In addition, 30D8 inhibited 50% of laser-induced choroidal neovascularization (CNV) in mice. Humanized 30D8 (hu30D8) showed similar in vitro and in vivo activities as the parental hamster antibody. A crystal structure of the hu30D8 Fab/CXCL12α complex in combination with mutational analysis revealed a "hot spot" around residues Asn(44)/Asn(45) of CXCL12α and part of the RFFESH region required for CXCL12α binding to CXCR4 and CXCR7. Finally, hu30D8 exhibited fast clearance in cynomolgus monkeys but not in rats.
  
  
  CONCLUSIONS
  CXCL12 is an attractive target for treatment of cancer and inflammation-related diseases; hu30D8 is suitable for testing this hypothesis in humans.
authors.affiliation: Genentech, Inc., South San Francisco, California, USA.
authors.forename: Cuiling
authors.initials: C
authors.lastname: Zhong
authors.forename: Jianyong
authors.initials: J
authors.lastname: Wang
authors.forename: Bing
authors.initials: B
authors.lastname: Li
authors.forename: Hong
authors.initials: H
authors.lastname: Xiang
authors.forename: Mark
authors.initials: M
authors.lastname: Ultsch
authors.forename: Mary
authors.initials: M
authors.lastname: Coons
authors.forename: Terence
authors.initials: T
authors.lastname: Wong
authors.forename: Nancy Y
authors.initials: NY
authors.lastname: Chiang
authors.forename: Suzy
authors.initials: S
authors.lastname: Clark
authors.forename: Robyn
authors.initials: R
authors.lastname: Clark
authors.forename: Leah
authors.initials: L
authors.lastname: Quintana
authors.forename: Peter
authors.initials: P
authors.lastname: Gribling
authors.forename: Eric
authors.initials: E
authors.lastname: Suto
authors.forename: Kai
authors.initials: K
authors.lastname: Barck
authors.forename: Racquel
authors.initials: R
authors.lastname: Corpuz
authors.forename: Jenny
authors.initials: J
authors.lastname: Yao
authors.forename: Rashi
authors.initials: R
authors.lastname: Takkar
authors.forename: Wyne P
authors.initials: WP
authors.lastname: Lee
authors.forename: Lisa A
authors.initials: LA
authors.lastname: Damico-Beyer
authors.forename: Richard D
authors.initials: RD
authors.lastname: Carano
authors.forename: Camellia
authors.initials: C
authors.lastname: Adams
authors.forename: Robert F
authors.initials: RF
authors.lastname: Kelley
authors.forename: Weiru
authors.initials: W
authors.lastname: Wang
authors.forename: Napoleone
authors.initials: N
authors.lastname: Ferrara
mesh_terms: D020533:Angiogenesis Inhibitors / Q000008:administration & dosage / Q000737:chemistry / Q000494:pharmacology; D000818:Animals; D000893:Anti-Inflammatory Agents / Q000008:administration & dosage / Q000737:chemistry / Q000494:pharmacology; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D061067:Antibodies, Monoclonal, Humanized / Q000008:administration & dosage / Q000737:chemistry / Q000494:pharmacology*; D000970:Antineoplastic Agents / Q000008:administration & dosage / Q000737:chemistry / Q000494:pharmacology*; D001169:Arthritis, Experimental / Q000188:drug therapy / Q000378:metabolism; D045744:Cell Line, Tumor; D054377:Chemokine CXCL12 / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism; D006224:Cricetinae; D004195:Disease Models, Animal; D004353:Drug Evaluation, Preclinical; D004357:Drug Synergism; D018604:Epitope Mapping; D005260:Female; D006801:Humans; D051379:Mice; D008958:Models, Molecular; D009362:Neoplasm Metastasis; D009369:Neoplasms / Q000188:drug therapy / Q000473:pathology; D011487:Protein Conformation; D047368:Tumor Burden / Q000187:drug effects; D042461:Vascular Endothelial Growth Factor A / Q000037:antagonists & inhibitors; D023041:Xenograft Model Antitumor Assays
chemical_list: D020533:Angiogenesis Inhibitors; D000893:Anti-Inflammatory Agents; D000911:Antibodies, Monoclonal; D061067:Antibodies, Monoclonal, Humanized; D000970:Antineoplastic Agents; D054377:Chemokine CXCL12; D042461:Vascular Endothelial Growth Factor A